Multiplex strand displacement amplification (SDA) and detection of DNA sequences from Mycobacterium tuberculosis and other mycobacteria

Strand Displacement Amplification (SDA) is an isothermal, in vitro method of amplifying a DNA target sequence prior to detection [Walker et al (1992) Nucleic Acids Res., 20, 1691 – 1693]. Here we describe a multiplex form of SDA that allows two target sequences and an internal amplification control...

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Bibliographic Details
Published in:Nucleic acids research 1994-07, Vol.22 (13), p.2670-2677
Main Authors: Walker, G.Terrance, Nadeau, James G., Spears, Patricia A., Schram, James L., Nycz, Colleen M., Shank, Daryl D.
Format: Article
Language:English
Subjects:
Base Sequence
DNA, Bacterial - analysis
Molecular Sequence Data
Mycobacterium - genetics
Mycobacterium tuberculosis
Mycobacterium tuberculosis - genetics
Nucleic Acid Amplification Techniques
Regulatory Sequences, Nucleic Acid
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Summary:Strand Displacement Amplification (SDA) is an isothermal, in vitro method of amplifying a DNA target sequence prior to detection [Walker et al (1992) Nucleic Acids Res., 20, 1691 – 1693]. Here we describe a multiplex form of SDA that allows two target sequences and an internal amplification control to be co-amplified by a single pair of primers after common priming sequences are spontaneously appended to the ends of target fragments. Multiplex SDA operates at a single temperature, under the same simple protocol previously developed for single-target SDA. We applied multiplex SDA to co-amplification of a target sequence (IS6110) that is specific to members of the Mycobacterium tuberculosis-complex and a target (16S ribosomal gene) that is common to most clinically relevant species of mycobacteria. Both targets are amplified 108-fold during a 2 hour, single temperature incubation. The relative sensitivity of the system was evaluated across a number of clinically relevant mycobacteria and checked for crossreactivity against organisms that are closely related to mycobacteria.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/22.13.2670